Martin Navrátil

Effects of different excitation and detection spectral regions on room temperature chlorophyll a fluorescence parameters

The effects of different spectral region of excitation and detection of chlorophyll (Chl) a fluorescence at room temperature on the estimation of excitation energy utilization within photosystem (PS) 2 were studied in wild-type barley (Hordeum vulgare L. cv. Bonus) and its Chl b-less mutant chlorina f2 grown under low and high irradiances [100 and 1 000 µmol(photon) m−2 s−1]. Three measuring spectral regimes were applied using a PAM 101 fluorometer: (1) excitation in the red region (maximum at the wavelength of 649 nm) and detection in the far-red region beyond 710 nm, (2) excitation in the blue region (maximum at the wavelength of 461 nm) and detection beyond 710 nm, and (3) excitation in the blue region and detection in the red region (660– 710 nm). Non-photochemical quenching of maximal (NPQ) and minimal fluorescence (SV0), determined by detecting Chl a fluorescence beyond 710 nm, were significantly higher for blue excitation as compared to red excitation. We suggest that this results from higher non-radiative dissipation of absorbed excitation energy within light-harvesting complexes of PS2 (LHC2) due to preferential excitation of LHC2 by blue radiation and from the lower contribution of PS1 emission to the detected fluorescence in the case of blue excitation. Detection of Chl a fluorescence originating preferentially from PS2 (i.e. in the range of 660–710 nm) led to pronounced increase of NPQ, SV0, and the PS2 photochemical efficiencies (FV/FM and FV′/FM′), indicating considerable underestimation of these parameters using the standard set-up of PAM 101. Hence PS1 contribution to the minimal fluorescence level in the irradiance-adapted state may reach up to about 80 %.

The development of antenna complexes of barley (Hordeum vulgare cv. Akcent) under different light conditions as judged from the analysis of 77 K chlorophyll a fluorescence spectra.

Using 77 K chlorophyll a (Chl a) fluorescence spectra in vivo, the development was studied of Photosystems II (PS II) and I (PS I) during greening of barley under intermittent light followed by continuous light at low (LI, 50 μmol m−2 s−1) and high (HI, 1000 μmol m−2 s−1) irradiances. The greening at HI intermittent light was accompanied with significantly reduced fluorescence intensity from Chl b excitation for both PS II (F685) and PS I (F743), in comparison with LI plants, indicating that assembly of light-harvesting complexes (LHC) of both photosystems was affected to a similar degree. During greening at continuous HI, a slower increase of emission from Chl b excitation in PS II as compared with PS I was observed, indicating a preferred reduction in the accumulation of LHC II. The following characteristics of 77 K Chl a fluorescence spectra documented the photoprotective function of an elevated content of carotenoids in HI leaves: (1) a pronounced suppression of Soret region of excitation spectra (410–450 nm) in comparison with the red region (670–690 nm) during the early stage of greening indicated a strongly reduced excitation energy transfer from carotenoids to the Chl a fluorescing forms within PS I and PS II; (2) changes in the shape of the excitation band of Chl b and carotenoids (460–490 nm) during greening under continuous light confirmed that the energy transfer from carotenoids to Chl a within PS II remained lower as compared with the LI plants.

Blue radiation stimulates photosynthetic induction in Fagus sylvatica L.

This study was designed to test the hypothesis that the spectral composition of incident radiation, as defined by the relative proportions of blue (B; λmax = 455 nm) and red (R; λmax = 625 nm) photons, can affect photosynthetic induction, since B photons stimulate stomatal opening and are more effectively absorbed by leaves than R photons. Different stages of photosynthetic induction, primarily determined by the photo-modulation of Rubisco activity and stomata opening, were investigated in dark-adapted leaves of Fagus sylvatica transferred to saturating irradiance [800µmol(photon) m−2 s−1] at B/R ratios of 1/3, 1/1, or 3/1.In agreement with our hypothesis, photosynthesis was induced faster by irradiance with a high B/R ratio (3/1); as demontrated by a higher IS60 (induction state 60 s after leaf illumination) and lower T90 (the time period required to reach 90 % of maximum steady-state photosynthesis). However, there were no differences in induction between leaves receiving equal (1/1) and low (1/3) B/R ratios. Electron transport was highly sensitive to radiation quality, exhibiting faster induction kinetics with increasing B/R ratio. Such stimulation of carbon-assimilatory processes corresponds with faster activation of Rubisco and lower non-photochemical quenching (NPQ) as the proportion of B photons is increased. In contrast, the kinetics of stomatal opening was independent of the spectral composition of incoming radiation. Since slightly higher absorption efficiency of high B/R radiation does not fully explain the changes in induction kinetics, the other possible mechanisms contributing to the stimulation of electron transport and Rubisco activity are discussed.

Assessing the consistency of optical properties measured in four integrating spheres

ABSTRACT Leaf optical properties (LOPs) determine the radiation regime of vegetation, thus being key input parameters in leaf and canopy radiative transfer models. It is of great importance to know the uncertainties originating from the LOP measurements. The most common approach to measure LOP uses integrating spheres. They allow measurements of both directional-hemispherical reflectance factor (R) and transmittance factor (T). However, sphere’s design, inner surface coating and measurement protocol differ among the spheres’ models and manufacturers. Our main goal was to evaluate the consistency of optical properties measured in four integrating spheres (Dualsphere, Labspehre, ASD, and Li-cor). Our test samples were three Spectralon® panels, four artificial materials and leaves from six common broadleaf tree species. Results showed that spectra measured in the four integrating spheres were generally similar in the spectral region between 400 and 1600 nm. The average standard deviation computed among the spectra of all samples measured in all spheres was around 0.023 (and varied between 0.005 and 0.044). Statistically significant differences were detected mainly between Dualsphere and Li-cor spheres.

Relation of chlorophyll fluorescence sensitive reflectance ratios to carbon flux measurements of montanne grassland and norway spruce forest ecosystems in the temperate zone.

We explored ability of reflectance vegetation indexes (VIs) related to chlorophyll fluorescence emission (R 686/R 630, R 740/R 800) and de-epoxidation state of xanthophyll cycle pigments (PRI, calculated as (R 531 − R 570)/(R 531 − R 570)) to track changes in the CO2 assimilation rate and Light Use Efficiency (LUE) in montane grassland and Norway spruce forest ecosystems, both at leaf and also canopy level. VIs were measured at two research plots using a ground-based high spatial/spectral resolution imaging spectroscopy technique. No significant relationship between VIs and leaf light-saturated CO2 assimilation (A MAX) was detected in instantaneous measurements of grassland under steady-state irradiance conditions. Once the temporal dimension and daily irradiance variation were included into the experimental setup, statistically significant changes in VIs related to tested physiological parameters were revealed. ΔPRI and Δ(R 686/R 630) of grassland plant leaves under dark-to-full sunlight transition in the scale of minutes were significantly related to A MAX (R 2 = 0.51). In the daily course, the variation of VIs measured in one-hour intervals correlated well with the variation of Gross Primary Production (GPP), Net Ecosystem Exchange (NEE), and LUE estimated via the eddy-covariance flux tower. Statistical results were weaker in the case of the grassland ecosystem, with the strongest statistical relation of the index R 686/R 630 with NEE and GPP.

Partitioning of absorbed light energy within photosystem II in barley can be affected by chloroplast movement

Plants have developed many ways to protect reaction centres of photosystems against overexcitation. One of the mechanisms involves reduction of the leaf absorption cross-section by light-induced chloroplast avoidance reaction. Decrease in the probability of photon absorption by the pigments bound within photosystem II (PSII) complexes leads to the increase in quantum yield of PSII photochemistry (ΦPSII). On the other hand, the decrease of PSII excitation probability causes reduction of chlorophyll a fluorescence intensity which is manifested as the apparent increase of determined quantum yield of regulated light-induced non-photochemical quenching (ΦNPQ). Absorption of different light intensity by phototropins led to the different chloroplast distribution within barley leaves, estimated by measurement of the leaf transmittance. Due to a weak blue light used for transmittance measurements, leaves exposed to actinic light with wavelengths longer than 520 nm undergo chloroplast accumulation reaction, in contrast with leaves exposed to light with shorter wavelengths, that showed a different extent of chloroplast avoidance reaction. Based on the ΦNPQ action spectra measured simultaneously with the transmittance, the influence of different chloroplast distribution on ΦNPQ was assessed. The analysis of results showed that decrease in the leaf absorption cross-section due to increasing part of chloroplasts reaching profile position significantly affected the partitioning of excitation energy within PSII and such rearrangement also distorted measured ΦNPQ and cannot be neglected in its interpretation. When the majority of chloroplasts reached profile position, the photoprotective effect appeared to be the most prominent for strong blue light that has the highest absorption in the upper leaf layers in comparison with green or red ones.